【摘要】 目的建立一种简便、实用和适合于免疫放射分析的抗体固化技术。方法制备羰基二咪唑（ＣＤＩ）活化的聚苯乙烯珠，提纯兔抗人促甲状腺素（ｈＴＳＨ）抗血清ＩｇＧ，选择ＩｇＧ最佳稀释度，使其与ＣＤＩ活化珠共价交联，制成固相抗体。测定固相抗体与ｈＴＳＨ－１２５Ⅰ－抗ｈＴＳＨ单克隆抗体的结合率及其重复性和稳定性，将其应用于促甲状腺素免疫放射分析，并与对照试剂盒作比较。结果ＩｇＧ（１ｍｇ／ｍｌ）的最佳稀释度为１６４（１５μｇ／ｍｌ），与ＣＤＩ活化珠结合牢固，经８次洗涤未见脱落。固相抗体与ｈＴＳＨ－１２５Ⅰ－抗ｈＴＳＨ单克隆抗体的结合率与ｈＴＳＨ浓度成正相关。批内和批间变异系数分别为２．５％～３．１％（ｎ＝１０）和２．９％～４．１％（ｎ＝１８）。非特异结合率为０．１２％±０．０４％（ｎ＝５０）。固相抗体应用于促甲状腺素免疫放射分析的各项质控指标与对照试剂盒一致，血清标本的ｈＴＳＨ测定值和对照试剂盒所测值的相关系数（ｒ）＝０．９８６，Ｐ＜０．０１（ｎ＝８６）。结论羰基二咪唑固化抗体是一种简便实用、重复性和稳定性好、适合于免疫放射分析的抗体固化技术更多还原

【Abstract】 Objective To establish a method for immobilization of antibodies which is simple, practical and suitable for immunoradiometric assay (IRMA). Methods Polystyrene beads were activated with carbonyldiimidazole (CDI). IgG was purified from rabbit antihuman thyrotropin (hTSH) antiserum. Optimal concentration of IgG was selected. Immobilized antibodies were prepared by covalently coupling IgG to CDIactivated polystyrene beads. Bindings of immobilized antibodies and hTSH125Ⅰanti hTSH McAb, its reproducibility and stability were determined. These immobilized antibodies were applied to hTSH IRMA. The results of IRMA were compared with those of control kits. Results The optimal concentration of IgG (1 mg/ml) was 1∶64 (15 μg/ml).Linkage of IgG and CDIactivated polystyrene beads was very firm. Bindings of immobilized antibodies and hTSH125ⅠMcAb were directly proportional to the concentration of hTSH. CVs of inter and intra tests were 2.5%～3.1% (n=10) and 2.9%～4.1% (n=18) respectively. None specific binding was 0.12%±0.04% (n=50). Quality characteristics of hTSH IRMA, which CDIactivated beads coupled with IgG were applied to, accorded with those of control kits. The concentration of hTSH of serum samples indicated a good correlation between our method and control kits (r=0.986,P<0.01,n=86). Conclusions Immobilization of antibodies using CDI is a simple, practical, reproducible and stable method for preparing immobilized antibodies with evidence of utility for hTSH IRMA.更多还原